Selection and application of broad-specificity human domain antibody for simultaneous detection of Bt Cry toxins
文献类型： 外文期刊
作者： Xu, Chongxin 1 ; Zhang, Xiao 1 ; Liu, Xiaoqin; Liu, Yuan 1 ; Hu, Xiaodan 1 ; Zhong, Jianfeng 1 ; Zhang, Cunzheng 1 ; L 1 ;
作者机构： 1.Jiangsu Acad Agr Sci, Inst Food Qual Safety & Detect, State Key Lab Breeding Base, Key Lab Food Qual & Safety Jiangsu Prov, Nanjing 210014, Jiangsu, Peoples R China
2.Jiangsu Acad Agr Sci, Inst F
关键词： Bt Cry toxins;Human domain antibody;ELISA;Soluble expression
期刊名称：ANALYTICAL BIOCHEMISTRY （ 影响因子：3.365； 五年影响因子：3.049 ）
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收录情况： SCI
摘要： Bt Cry toxin is a kind of bio-toxins that used for genetically modified crops (GMC) transformation widely. In this study, total 15 positive clones could bind the Bt Cry toxins which isolated from a human domain antibody library by 5 rounds affinity selection. According to analyzing of PCR amplification and enzyme linked immunosorbent assay (ELISA), the most positive phage domain antibody (named F5) gene was cloned into the pET26b vector and expressed in E. coli BL21. The purified antibody was used to develop an indirect competitive ELISA (IC-ELISA) for Cry1Ab, Cry1Ac, Cry1B, Cry1C and Cry1F toxins, respectively. The working range of detection for standard curves in IC-ELISA were 0.258-1.407 mu g/mL, the medium inhibition concentration (IC50) were 0.727-0.892 mu g/mL and detection limit (IC10) were 0.029-0.07414 mu g/mL for those Bt Cry toxins. The affinity of F5 domain antibody with Cry1Ab, Cry1Ac, Cry1B, Cry1C and Cry1F toxins were 1.21-5.94 x 10(7) M-1. The average recoveries of the 5 kinds of Bt Cry toxins from spiked wheat samples were ranged from 81.2%-100.8% with a CV at 2.5%-9.4%. The results showed that we successfully obtained the broad-specificity human domain antibody for simultaneous detection of Bt Cry toxins in agricultural product samples. (C) 2016 Elsevier Inc. All rights reserved.
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